Overview | PrinterFriendlyVersion |
Ex/Em(nm) | 384/540 |
MW | ~10K |
CAS# | N/A |
Solvent | Water |
Storage | F/D/L |
Category | CellBIOLOGy pHandIonIndicators |
Related | CellMetabolism |
Spectrum | AdvancedSpectrumViewer |
1.Preparecellsasdesired.Forexample,plateadherentcellsovernightingrowthmediumat40,000to80,000cells/well/100µLfor
96-wellor10,000to20,000cells/well/25µLfor384-wellplates.
Note: Eachcelllineshouldbeevaluatedonanindividualbasistodeterminetheoptimalcelldensity.
2.PrepareRatioWorks™PDMPODextranloadingsolution:
2.1 Preparea1mg/mLstocksolutionofRatioWorks™PDMPODextranin1mLofsterilewaterorHanksand20mMHepesbuffer(HHBS).Thestocksolutionshouldbeusedpromptly.Anyunusedsolutionneedtobealiquotedandrefrozenat<-20oC.
Note:Avoidrepeatedfreeze-thawcycles,andprotectfromlight.
2.2 Preparea20-100ug/mLRatioWorks™PDMPODextranloadingsolutioninHHBS.
3. RunEndocytosisAssay
3.1 Removethemedium,andadd100µL/well(96-wellplate)or25µL/well(384-wellplate)RatioWorks™PDMPODextranloadingsolutionintothecellplate(fromStep2.2).
Note1: ItisimportanttoreplacethegrowthmediumwithHHBSbuffer(100µL/wellfor96-wellplateor25µL/wellfor384-wellplatebeforedye-loading)ifyourcompoundsinterferewiththeserum.
Note2:RapidtraffickingofRatioWorks™PDMPOdextranfromearlyendosomestolateendosomesandsubsequentfusionwithlysosomescanoccur.ToaidthevisualizationofRatioWorks™PDMPOdextranwithintheendosomes,werecommendincreasingthelabelingconcentrationanddecreasingtheloadingtime,andimagingimmediately.
3.2 Incubatethedye-loadingplateatcellincubatorfor5to20minutes.
3.3 Washandreplacethedye-loadingsolutionwithHHBSorgrowthmedium.
3.4 RuntheendocytosisassaybymonitoringthefluorescenceatEx=360nm,andEm=450and540nmforratiomeasurements.
References&Citations | CitationExplorer |
iTRAQ-basedproteomicanalysisofthemetabolismmechanismassociatedwithsiliconresponseinthemarinediatomThalassiosirapseudonana
Authors:ChaoDu,Jun-RongLiang,Dan-DanChen,BinXu,Wen-HaoZhuo,Ya-HuiGao,Chang-PingChen,ChrisBowler,WenZhang
Journal:Journalofproteomeresearch(2014):720--734